LOCUS       sequence-60644-e        6152 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6152)
  AUTHORS   Roczniak-Ferguson A, Petit CS, Froehlich F, Qian S, Ky J, Angarola 
            B, Walther TC, Ferguson SM
  TITLE     The Transcription Factor TFEB Links mTORC1 Signaling to 
            Transcriptional Control of Lysosome Homeostasis.
  JOURNAL   Sci Signal. 2012 Jun 12;5(228):ra42.
  PUBMED    22692423
REFERENCE   2  (bases 1 to 6152)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci 
            Signal."; date: "2012-06-12"; volume: "5(228)"; pages: "ra42"
COMMENT     Sequence Label: pegfp-N1-TFEB-S3A,R4A
FEATURES             Location/Qualifiers
     source          1..6152
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      628..637
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      632..641
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2092..2101
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2098..2100,2101..2103,2104..2817)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2098..2100,2101..2103,2104..2814)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2143..2164)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2404..2423)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2751..2772
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2940..3061
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2977..2996)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3031..3050
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3068..3523)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3205..3226)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3417..3436
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3550..3654
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3656..4013
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3656..3847
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3679..3699)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3864..3999
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4048..4842
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4048..4842
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4102..4121)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4712..4731
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5030..5049)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5074..5121
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5450..6038
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5939..5958
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttgcc accatggcgg cagccatagg gttgcgcatg
      661 cagctcatgc gggagcaggc gcagcaggag gagcaacggg agcgcatgca gcaacaggct
      721 gtcatgcatt acatgcagca gcagcagcag cagcaacagc agcagctcgg agggccgccc
      781 accccggcca tcaatacccc cgtccacttc cagtcgccac cacctgtgcc tggggaggtg
      841 ttgaaggtgc agtcctacct ggagaatccc acatcctacc atctgcagca gtcgcagcat
      901 cagaaggtgc gggagtacct gtccgagacc tatgggaaca agtttgctgc ccacatcagc
      961 ccagcccagg gctctccgaa acccccacca gccgcctccc caggggtgcg agctggacac
     1021 gtgctgtcct cctccgctgg caacagtgct cccaatagcc ccatggccat gctgcacatt
     1081 ggctccaacc ctgagaggga gttggatgat gtcattgaca acattatgcg tctggacgat
     1141 gtccttggct acatcaatcc tgaaatgcag atgcccaaca cgctacccct gtccagcagc
     1201 cacctgaatg tgtacagcag cgacccccag gtcacagcct ccctggtggg cgtcaccagc
     1261 agctcctgcc ctgcggacct gacccagaag cgagagctca cagatgctga gagcagggcc
     1321 ctggccaagg agcggcagaa gaaagacaat cacaacttaa ttgaaaggag acgaaggttc
     1381 aacatcaatg accgcatcaa ggagttggga atgctgatcc ccaaggccaa tgacctggac
     1441 gtgcgctgga acaagggcac catcctcaag gcctctgtgg attacatccg gaggatgcag
     1501 aaggacctgc aaaagtccag ggagctggag aaccactctc gccgcctgga gatgaccaac
     1561 aagcagctct ggctccgtat ccaggagctg gagatgcagg ctcgagtgca cggcctccct
     1621 accacctccc cgtccggcat gaacatggct gagctggccc agcaggtggt gaagcaggag
     1681 ctgcctagcg aagagggccc aggggaggcc ctgatgctgg gggctgaggt ccctgaccct
     1741 gagccactgc cagctctgcc cccgcaagcc ccgctgcccc tgcccaccca gccaccatcc
     1801 ccattccatc acctggactt cagccacagc ctgagctttg ggggcaggga ggacgagggt
     1861 cccccgggct accccgaacc cctggcgccg gggcatggct ccccattccc cagcctgtcc
     1921 aagaaggatc tggacctcat gctcctggac gactcactgc taccgctggc ctctgatcca
     1981 cttctgtcca ccatgtcccc cgaggcctcc aaggccagca gccgccggag cagcttcagc
     2041 atggaggagg gcgatgtgct gaaggtaccg cgggcccggg atccaccggt cgccaccatg
     2101 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     2161 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     2221 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     2281 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     2341 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     2401 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     2461 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     2521 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     2581 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     2641 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     2701 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     2761 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc
     2821 ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     2881 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     2941 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3001 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3061 aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     3121 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     3181 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     3241 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     3301 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     3361 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     3421 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     3481 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     3541 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3601 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
     3661 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3721 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3781 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3841 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3901 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     3961 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
     4021 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4081 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4141 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4201 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
     4261 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4321 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4381 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4441 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4501 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4561 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4621 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4681 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4741 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4801 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4861 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     4921 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4981 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga
     5041 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     5101 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     5161 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     5221 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     5281 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5341 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5401 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5461 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5521 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5581 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5641 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5701 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5761 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5821 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5881 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5941 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6001 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6061 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     6121 attctgtgga taaccgtatt accgccatgc at
//
